CC12 Induces Apoptotic Cell Death and Cell Cycle Arrest in Human Glioblastoma Cell Lines and Mouse Xenograft Model.

Among central nervous system tumors, glioblastoma (GBM) is the most common and the most malignant type. Even under current standard treatments, the overall survival rate is still low and the recurrence rate is high. Therefore, developing novel and effective therapy is urgently needed. CC12, a synthesized small molecule, was evaluated for the potential anti-GBM effects in two GBM cell lines, U87MG and U118MG. The observations of cell morphology, MTT assay, flow cytometry-based apoptosis after CC12 treatment, were conducted. Western blot was performed for the investigation of the apoptotic mechanism. Positron emission tomography scan analysis and bioluminescent imaging assay using a mouse xenograft model were performed for the effect of CC12 in vivo. After treated by 10 µM CC12 for 24 h, both U118MG and U87MG cells showed tumor cell death. MTT assay results showed that the survival rates decreased when the CC12 concentrations or the treatment periods increased. Ki-67 expression and flow cytometry results indicated that the proliferation was inhibited in GBM cells, and G1 phase arrest was shown. The results of 7-AAD, Br-dUTP, and JC-1 staining all showed the apoptosis of GBM cells after CC12 treatment. Increased γH2AX, caspase-3, and poly (ADP-ribose) polymerase (PARP) levels meant the DNA damage, and increased Bcl2 family proteins after CC12 treatment indicated the intrinsic apoptotic pathway was involved in CC12 induced apoptosis. Furthermore, CC12 can induce the decrease of tumor prognostic marker DcR3. In vivo experiment results showed the effect of CC12 on tumor size reduction of CC12. In addition, the ability to cross the brain-blood barrier of CC12 was also confirmed. CC12 may have anti-tumor ability through the regulation of cell cycle and apoptosis in vitro and in vivo.

A Synergistic Anti-Cancer Effect of Troglitazone and Lovastatin in a Human Anaplastic Thyroid Cancer Cell Line and in a Mouse Xenograft Model.

Anaplastic thyroid cancer (ATC) is a malignant subtype of thyroid cancers and its mechanism of development remains inconclusive. Importantly, there is no effective strategy for treatment since ATC is not responsive to conventional therapies, including radioactive iodine therapy and thyroid-stimulating hormone suppression. Here, we report that a combinational approach consisting of drugs designed for targeting lipid metabolism, lovastatin (an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, HMGCR) and troglitazone (an agonist of peroxisome proliferator-activated receptor gamma, PPARγ), exhibits anti-proliferation in cell culture systems and leads to tumor regression in a mouse xenograft model. The composition contains a sub-lethal concentration of both drugs and exhibits low toxicity to certain types of normal cells. Our results support a hypothesis that the inhibitory effect of the combination is partly through a cell cycle arrest at G0/G1 phase, as evidenced by the induction of cyclin-dependent kinase inhibitors, p21cip and p27kip, and the reduction of hyperphosphorylated retinoblastoma protein (pp-Rb)-E2F1 signaling. Therefore, targeting two pathways involved in lipid metabolism may provide a new direction for treating ATC.

A Randomized, Double-Blind Clinical Study on Blood Pressure Reduction and Blood Lipid Profile Amelioration on Treatment with Ankascin 568.

Hypertension and cardiovascular complications are the leading causes of death worldwide. Antihypertensive drugs often cause various side effects, and improper use of antihypertensive medications can result in irreparable damage. Edible fungi of the Monascus species have been used as traditional Chinese medicines in Southeast Asia for several centuries. The fermented products of Monascus purpureus NTU 568 (ANKASCIN 568) possess a number of functional secondary metabolites including the anti-inflammatory pigments monascin (MS) and ankaflavin (AK). In this study, a double-blind, placebo-controlled clinical trial was performed in which patients with mild to moderate hypertension were randomly assigned to receive placebo or two 500-mg capsules of Ankascin 568 for 8 weeks. The effects of this treatment on the regulation of blood pressure (BP) were then examined. The results showed that systolic blood pressure (SBP) decreased from 141.6 ± 12.0 to 133.9 ± 14.4 mmHg (P < 0.05), and diastolic blood pressure (DBP) decreased from 91.7 ± 8.1 to 84.8 ± 7.4 mmHg (P < 0.05). Moreover, Ankascin 568 treatment effectively reduced serum triglycerides and total cholesterol (TC), increased high-density lipoprotein cholesterol (HDL-C), and reduced low-density lipoprotein cholesterol (LDL-C) levels, thereby improving the serum lipid profile. Additionally, administration of Ankascin 568 did not cause significant rhabdomyolysis nor impaired the metabolic or physiological functions of the liver or kidney. In conclusion, patients administered Ankascin 568 for 8 weeks exhibited significant in reduction of SBP, serum TC and LDL-C levels, which should contribute to better cardiovascular health.

Simvastatin Inhibits Cell Proliferation and Migration in Human Anaplastic Thyroid Cancer.

Malignant human anaplastic thyroid cancer (ATC) is pertinacious to conventional therapies. The present study investigated the anti-cancer activity of simvastatin and its underlying regulatory mechanism in cultured ATC cells. Simvastatin (0-20 µM) concentration-dependently reduced cell viability and relative colony formation. Depletions of mevalonate (MEV) and geranylgeranyl pyrophosphate (GGpp) by simvastatin induced G1 arrest and increased apoptotic cell populations at the sub-G1 phase. Adding MEV and GGpp prevented the simvastatin-inhibited cell proliferation. Immunoblotting analysis illustrated that simvastatin diminished the activation of RhoA and Rac1 protein, and this effect was prevented by pre-treatment with MEV and GGpp. Simvastatin increased the levels of p21cip and p27kip proteins and reduced the levels of hyperphosphorylated-Rb, E2F1 and CCND1 proteins. Adding GGpp abolished the simvastatin-increased levels of p27kip protein, and the GGpp-caused effect was abolished by Skp2 inhibition. Introduction of Cyr61 siRNA into ATC cells prevented the epidermal growth factor (EGF)-enhanced cell migration. The EGF-induced increases of Cyr61 protein expression and cell migration were prevented by simvastatin. Taken together, these results suggest that simvastatin induced ATC proliferation inhibition through the deactivation of RhoA/Rac1 protein and overexpression of p21cip and p27kip, and migration inhibition through the abrogation of Cyr61 protein expression.

Association between Anemia and Auditory Threshold Shifts in the US Population: National Health and Nutrition Examination Survey.

Existing evidence indicates that both iron deficiency anemia and sickle cell anemia have been previously associated with hearing loss. However, human data investigating the association between anemia and auditory threshold shifts at different frequencies in the adolescent, adult and elderly population are extremely limited to date. Therefore, this cross-sectional study used the dataset from the US National Health and Nutrition Examination Survey from 2005 to 2012 to explore differences in low- or high-frequency hearing thresholds and hearing loss prevalence between participants with and without anemia. A total of 918 patients with anemia and 8213 without anemia were included. Results indicated that low- and high-frequency pure tone average were significantly higher in patients with anemia than that in those without anemia in the elderly, but not in adult or adolescent population. In addition, the prevalence of low-frequency hearing loss but not high-frequency hearing loss was also higher in patients with anemia than in those without anemia in the elderly population. After adjusting various confounders, multiple regression models still indicated that patients with anemia tended to have larger threshold shift. In conclusion, anemia was associated with auditory threshold shifts in the elderly population, especially those vulnerable to low-frequency hearing loss.

Outcome of untethering for symptomatic spina bifida occulta with lumbosacral spinal cord tethering in 31 patients: analysis of preoperative prognostic factors.

BACKGROUND CONTEXT: The most important goal for treating symptomatic lumbosacral spinal cord tethering is early untethering. PURPOSE: To investigate preoperative symptoms that may have affected the outcome. STUDY DESIGN: Patients with or without improvement and with or without favorable outcome after untethering were compared retrospectively by chart and image review. PATIENT SAMPLE: Thirty-one patients (age between 2 days to 25 years) with spina bifida occulta and symptomatic cord tethering were analyzed. Presenting symptoms (neurological deficits, urological dysfunction, and lower limb deformities) were assessed before and after untethering. OUTCOME MEASURES: Favorable outcome was defined as complete relief of symptoms or mild symptoms whereby patients are able to look after their own personal care without assistance. Unfavorable outcome was defined as moderate or severe disability whereby patients are unable to attend to their own bodily needs without assistance, are bedridden, or require constant nursing attention. METHODS: Differences in patient characteristics and presenting symptoms were compared between those with and without clinical improvement and favorable outcome. Multivariate logistic regression was used to identify prognostic factors affecting the outcome. RESULTS: The average age at surgery was 7.2 years, with a male-to-female ratio of 1.2. The average follow-up time was 4 years. At least one of the following symptoms was present in all patients: neurological deficits (83.9%), urological dysfunction (77.4%), or limb deformities (38.7%). After untethering, all patients had either symptoms stabilized (14 patients, 45.2%) or improved (17 patients, 54.8%), and 14 patients (45.2%) achieved total resolving of symptoms. Logistic regression confirmed that younger age (< or =2 years, odds ratio [OR] 22.0, p=.026), lipomas of filum terminale (OR 25.6, p=.042), and a poor anal tone (OR 10.4, p=.061) were positive prognostic factors for the improvement in symptoms. The functional outcome was determined by the age at surgery (OR 0.9 per year since 1 year old, p=.04) and the presence of limb deformities (OR 0.06, p=.017). CONCLUSIONS: In conclusion, our study suggests that untethering should be performed immediately once the patient shows evidence of symptomatic lumbosacral cord tethering, irrespective of age. Untethering can interrupt progression of symptoms, but sphincter dysfunction and muscle weakness are more likely to improve or resolve. Benefits can be seen in all patients, but young children (before 2 years old) have a higher chance to gain favorable outcome. Retethering is a main concern during follow-up, particularly for the more complicated lipomyelomeningoceles. Investigations using electrophysiologic and urodynamic studies are helpful for early detection of subtle symptomatic cord tethering or retethering.

Survival analysis of 459 adult patients with primary spinal cancer in England and Wales: a population-based study.

BACKGROUND: Primary spinal gliomas are rare. Because data from the Cancer Registry of England and Wales covered larger numbers of patients and longer durations of follow-up, our objective was to define prognostic factors for survival at a national population level. METHODS: From 1971 to 1995, data on 459 adult patients (age, >15 years) with primary spinal cancer from the Cancer Registry of England and Wales were analyzed. Median survival and CSRs with respect to 7 variables (age, sex, morphology, WHO grade, socioeconomic status, geographic region, and period of diagnosis) were calculated using the Kaplan-Meier method. Cox regression was performed for estimating HRs for death. RESULTS: The median survival and the 1-, 5-, and 10-year CSRs for the population were 10.2 years, 78.7%, 59.7%, and 50.5%, respectively. Univariate analysis revealed that age at diagnosis, morphology, WHO grade, and period of diagnosis affected the CSR. Multivariate analysis demonstrated 3 factors influencing survival: older age (ie, >60 years; HR = 2.67; P < .001), non-ependymomas (HR = 3.51; P < .001), and high-grade tumors (HR = 3.01; P < .001). The improved survival in recent periods was associated with an increased number of diagnosed ependymomas. CONCLUSION: This study identifies old age, non-ependymomas, and high-grade tumors as negative prognostic factors for patient survival. The results from this population-based study are very helpful for comparison with those of other hospital-based studies and for public health purposes.

Valproic acid inhibits glioblastoma multiforme cell growth via paraoxonase 2 expression.

We studied the potential mechanisms of valproic acid (VPA) in the treatment of glioblastoma multiforme (GBM). Using the human U87, GBM8401, and DBTRG-05MG GBM-derived cell lines, VPA at concentrations of 5 to 20 mM induced G2/M cell cycle arrest and increased the production of reactive oxygen species (ROS). Stress-related molecules such as paraoxonase 2 (PON2), cyclin B1, cdc2, and Bcl-xL were downregulated, but p27, p21 and Bim were upregulated by VPA treatment. VPA response element on the PON2 promoter was localized at position -400/-1. PON2 protein expression was increased in GBM cells compared with normal brain tissue and there was a negative correlation between the expression of PON2 and Bim. These findings were confirmed by the public Bredel GBM microarray (Gene Expression Omnibus accession: GSE2223) and the Cancer Genome Atlas GBM microarray datasets. Overexpression of PON2 in GBM cells significantly decreased intracellular ROS levels, and PON2 expression was decreased after VPA stimulation compared with controls. Bim expression was significantly induced by VPA in GBM cells with PON2 silencing. These observations were further shown in the subcutaneous GBM8401 cell xenograft of BALB/c nude mice. Our results suggest that VPA reduces PON2 expression in GBM cells, which in turn increases ROS production and induces Bim production that inhibits cancer progression via the PON2-Bim cascade.

Comparison of effects of socioeconomic and geographic variations on survival for adults and children with glioma.

OBJECT: The authors analyzed the effects of socioeconomic status (SES) and geographic variations on survival rates for adults and children with glioma, studying the data of 30,489 adults and 2940 children from the Cancer Registry in England and Wales. METHODS: The median survival time and crude survival rates for eight variables (age, sex, morphology, World Health Organization grade, tumor site, SES, geographic region, and period of diagnosis) were calculated using the Kaplan-Meier method. Distributions among different variables were compared using the chi-square test. Cox regressions were performed to estimate the hazard ratios (HR) to death. The median survival time and 1-, 5-, and 10-year crude survival rates for adults were 0.42 years and 29.1, 12, and 7.7%, respectively; the values for children were 9.33 years and 72.69, 54.32, and 49.5%, respectively. Similar gradients in SES from the south to the north exist in both populations (p < 0.001, chi-square test). Multivariate analyses revealed that all eight variables influenced survival in adults, including independent effects of sex (HR 0.94 for female, p < 0.001), SES (HR 1.03/quintile of deprivation, p < 0.001), and geographic region (HR 1.10 for outside southern England, p < 0.001). In children, only five of the eight variables affected survival; sex, SES, and geographic variation did not have an effect. CONCLUSIONS: Although age and tumor characteristics are well-known prognostic factors for both adults and children with glioma, SES and geographic variation also play significant roles in the survival of adults. The effects of SES and geographic variation may be directly related to the National Health Service in the United Kingdom.

Brain abscess in 142 patients: factors influencing outcome and mortality.

BACKGROUND: With the introduction of CT, stereotactic techniques, and broad-spectrum antibiotics, the outcome for brain abscess has dramatically improved. The purpose of this study was to identify prognostic factors by reviewing data on 142 patients with brain abscess. METHODS: Clinical data, including age, sex, medical history, duration of symptoms, initial neurological status, associated predisposing factors, laboratory data, treatment, and abscess characteristics, were considered as potential prognostic factors. A comparison was made between patients with favorable (GOS: moderate disability or good recovery) and those with unfavorable (GOS: death, persistent vegetative status, or severe disability) outcomes at discharge. Univariate (chi(2) analysis or Fisher's exact test) and multivariate logistic regression analyses were used to identify prognostic factors. Data were considered significant when the 2-tailed P value was lower than .05. RESULTS: There were 98 male and 44 female patients (male/female ratio, 2.2). Their average age at diagnosis was 41.5 years (range, 2-84 years). There were 105 patients with a favorable outcome and 37 with an unfavorable outcome. Both univariate and multivariate analyses indicated that patients who were male, had an initial GCS score >12, had no other septic complication, or had Gram-positive cocci grown in abscess cultures had better outcomes. No association was found between outcome and other factors, including age, focal neurological deficits, seizures, laboratory findings, characteristics of the abscesses, associated factors, and treatment modalities. CONCLUSIONS: With the advancement of imaging studies and broad-spectrum antibiotic therapies, the outcome of brain abscess depends on prompt awareness of the diagnosis and effective infection control.

Effects of socioeconomic and geographic variations on survival for adult glioma in England and Wales.

BACKGROUND: To investigate the effects of SES and geographic variations on survival for adult patients with glioma, we analyzed data from 30489 patients from the Cancer Registry in England and Wales. METHODS: Median survival and CSRs for 8 variables (age, sex, morphology, World Health Organization [WHO] grade, tumor site, SES, geographic regions, and periods of diagnosis) are calculated using the Kaplan-Meier method. Distributions among different variables are compared using chi(2) test. Cox regressions are performed for estimating HRs to death. RESULTS: The median survival and the 1-, 5-, and 10-year CSR in this population are 0.42 years, 29.1%, 12.0%, and 7.7%, respectively. There is a gradient in SES from the south to the north (chi(2) test, P < .001) and a gradual increment in higher SES from the early to the recent period (chi(2) test, P < .001). Mono- and multivariate analyses reveal that all the 8 variables influenced the survival (P < .05). Age (HR, 1.04 per year from 15 years, P < .001), WHO grade (1.21 per grade from grade I, P < .001), and morphology (HR from 1.23 to 1.89, compared with ependymoma, P < .05) are the most influential factors. However, there are also independent effects from SES (HR, 1.03 per quintile of deprivation, P < .001) and geographic regions (HR, 1.10 for outside southern England; P < .001) on the survival. CONCLUSIONS: Although age and tumor characteristics (morphology, WHO grade, tumor site) are well-known prognostic factors, SES and geographic variations also play a slight but significant role, and for more cost-effective allocation of health resources, alleviation on these 2 modifiable factors should be considered.

Survival analysis for adult glioma in England and Wales.

BACKGROUND AND PURPOSE: To investigate factors influencing survival for adult glioma at the national population level, data from the Cancer Registry in England and Wales were analyzed. Glioma was diagnosed in 32,267 patients during 1971-1990 and follow-up data were available for these patients until 1995. METHODS: Median survival and crude survival rates (CSR) for 8 variables (age, gender, International Classification of Diseases for Oncology [ICD-O] morphology, World Health Organization [WHO] grade, tumor site, deprivation, geographical region, and period of diagnosis) were calculated and tested using the Kaplan-Meier method and the log-rank test. Relative survival rates (RSR) were calculated using the life table of 1981. Cox multivariate regression was performed for estimating hazards ratios (HR) and tested using the log likelihood ratio test. RESULTS: The median survival and the 1-, 5-, and 10-year CSR for this population were 0.42 years, 29.1%, 12.0%, and 7.7%, respectively. The 1- and 5-year RSR were 29.6% and 12.3%, respectively. Survival was influenced by all 8 variables tested (p

Risk factors for chronic subdural hematoma after a minor head injury in the elderly: a population-based study.

Chronic subdural hematoma (CSDH) is one of the major comorbidities in elderly resulting in disability and death. Early recognition of CSDH is important for early management. However, manifestations of CSDH are nonspecific and subtle. Therefore, identification of risk factors of CSDH can offer clinical follow-up strategies for patients after episodes of head injury. The purpose of the study aimed at identifying risk factors of CSDH of Taiwanese. Analysis of data from the National Health Insurance provides important information on predictive factors influencing the early diagnosis of CSDH in elderly patients following minor head injuries. The current study is the first nationwide population-based study in Taiwan, showing that old age (≥75 years), male gender, and coexisting hydrocephalus are significantly predictive factors, irrespective to their medical comorbidities.

Survival analysis of children with primary malignant brain tumors in England and Wales: a population-based study.

Primary malignant brain tumor is the second most common cancer in children. To investigate factors affecting children's survival at a population level, data of 3,169 patients (age<15 years) from the Cancer Registry in England and Wales were used. They were diagnosed during 1971-1990 and followed up until 1995. Variables including age, gender, morphology, WHO grade, tumor site, socioeconomic status, geographical region, and period of diagnosis were available for analysis using the Kaplan-Meier method and the Cox hazards ratio (HR) regression. Results showed that the median survival and the 1-, 5-, and 10-year crude survival rate for this population were 8.7 years, 72.4, 54.0, and 49.2% respectively. Survival was influenced by age (HR 0.88/5 years), morphology (ependymoma HR 2.43), WHO grades (HR 1.42/grade), tumor sites (brain stem HR 2.11), and periods of diagnosis (HR 0.88/5 years). Gender, socioeconomic status, and geographical region did not affect their survival. Results from this population-based data are very helpful for comparison with other hospital-based studies and for public health purposes.

Survival analysis of 81 children with primary spinal gliomas: a population-based study.

Primary spinal gliomas are rare. Most clinical studies are based on single centers with small numbers of patients and limited length of follow-up. Because data from the Cancer Registry cover larger numbers of patients and longer durations of follow-up, our objective was to define prognostic factors that might predict the survival at a national population level. From 1971 to 1995, data of 81 children (age < 15 years) with primary spinal gliomas from the Cancer Registry of England and Wales were analyzed. Median survival and crude survival rates in respect of 7 variables (age, sex, morphology, WHO grade, socioeconomic status, geographical region, and period of diagnosis) were calculated using the Kaplan-Meier method. The Cox regression was performed for estimating hazard ratios (HR) for death. Results showed that the 1-, 5-, and 10-year crude survival rates for this population were 72.84, 60.49, and 58.0%, respectively. Both univariate and multivariate analyses revealed that only morphology (HR 2.79 for nonependymoma, p = 0.05) and WHO grade (HR 6.74 for high grade, p = 0.01) were significant prognostic factors. Results from this population-based study are very helpful for comparison with other population-based studies and for public health purposes.

Chronological changes on magnetic resonance images in a case of desmoplastic infantile ganglioglioma.

Desmoplastic infantile gangliogliomas (DIGs) are rare tumors during infancy. They often occur as huge cystic tumors in the frontal and parietal lobes, with their solid component being adjacent to the leptomeningeal membrane. This report presents a patient with DIG and intractable epilepsy. Due to hesitation by the patient's family in allowing surgery, this is the first time that the chronological changes in DIG have been observed on imaging studies. During the follow-up, the tumor changed from a pure solid tumor to a cystic one, which is a typical picture of DIG. Surgical pathology confirmed the diagnosis of DIG. However, for a patient with epilepsy, it is recommended that this condition should be treated immediately with surgery.